雄牛特异的SRY同源序列的扩增和分析

Amplification and Analysis of a Male-specific SRY Homologous Sequence of Cattle

利用人、兔、鼠SRY序列设计引物,应用PCR扩增牛的SRY序列,获得200bp的雄牛特异的扩增片段。克隆该扩增片段,获得重组质粒pCH21,进行序列分析,并与人、兔和鼠SRY的对应区域比较,具有高度同源性。用pCH21 DNA作探针与牛的基因组DNA酶切图谱杂交,显示了雄牛特异的1.7kb的杂交带。分析200bp的PCR扩增片段是牛的SRY基因片段。用同一对引物扩增人和山羊的DNA样品,也获得雄性特异的200bp的扩增片段。

Based on the highly ccnserved region of human, rabbit, mouse SRY (sex-determining region Y) sequences, the primers were designed for the amplification of the unknown homologous sequence of cattle by polymerase chain reaction (PCR). A 200bp male-specific amplifing fragment was obtained. The fragment shares high homology with the known SRY sequences and has been proposed to be a sequence of bovine SRY gene. By using the cloned fragment as a probe to hybridize to the Southern blot of Hind Ⅲ -digested cattle genomic DNA, a 1.7kb male-specific fragment was detected, and which is considered to contain the 200bp fragment. When the same pair of primers was used in amplification of human and goat DNA samples, the 200bp male-specific fragment was also obtained.